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S.C. Lin, J. Wong, P. Minasi, M. Muench; Isolation, in vitro Culture, and Characterization of Putative Fetal Trabecular Meshwork Cells . Invest. Ophthalmol. Vis. Sci. 2006;47(13):1879.
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The purpose of this study was to isolate and characterize fetal trabecular meshwork (FTM) cells for serial passage in culture. Cultured older adult TM cells often possess a restricted number of population doublings in the culture system, limiting the ability to perform expanded testing. FTM cells may proliferate more rapidly and abundantly, providing an excellent source of cells for the investigation of the molecular basis for primary open–angle glaucoma (POAG).
Institutional review board approval was obtained prior to tissue isolation. Fetal eyes from 24–week gestation abortions were procured and delicately dissected to isolate the developing TM tissue. Two primary cultures were achieved and passaged. Light microscopy was used to compare the putative FTM cells to previously cultured adult TM cells. Immunohistochemistry and western blot analysis was utilized to identify specific marker expression in the two cell types.
Both cell lines of putative FTM cells demonstrated similar microscopic characteristics as the adult TM cells, including monolayer formation, cobblestone pattern, and comparable size. FTM cells reached confluence after passaging in 3–4 days versus 7–10 days in the adult TM cells. Immunofluorescent staining was positive for actin, vimentin, fibronectin,aquaporin–1, and myocilin in both FTM and adult TM cells. Westerns showed substantial increase in myocilin after addition of 2000ng/L of dexamethasone.
Preliminary testing by microscopy and immunofluorescent markers suggest that FTM cells have properties that are similar to adult TM cells.Fetal tissues may be a useful source of abundant, rapidly dividing TM cells for in vitro investigation. The ability to do expanded research in this field may hasten the discovery of the molecular mechanisms for POAG development.
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